Sartorius_May21_NeutAntiMakingTheir - 14

Antibody Internalization

CD20
CD22

CD79b
CD3

CD71

CD19
mlgG1

100000.0
80000.0
60000.0
40000.0
20000.0
0

2
log(Concentration) μg/mL

CD20
CD22

CD79b
CD3

CD71

100000.0
80000.0
60000.0
40000.0
20000.0

3

0

2
log(Concentration) μg/mL

3

Median internalization (RL1-H) of Ramos

CD19
mlgG1

Median internalization (RL1-H) of Raji

Median internalization (RL1-H) of Jurkat

A.
CD19
mlgG1

CD20
CD22

CD79b
CD3

CD71

60000.0
50000.0
40000.0
30000.0
20000.0
10000.0
0

2
log(Concentration) μg/mL

B.
Jurkat

Raji

Ramos

mlgG1

CD71

CD3

CD20

CD19

CD22

CD79b

Figure 4: Serial dilution curves for internalization-labeled antibodies with a top concentration of 1 mg/mL in different cell types after
a three-hour incubation. Multiplexed positive and negative cell lines may be used in an ABI assay to generate high-content data in
one assay. Median fluorescent intensity (MFI) for Internalization Reagent-labeled antibodies after three hours. A serial dilution of
each antibody with a top concentration of 1 mg/mL was prepared and incubated with encoded Jurkat, Raji, and Ramos cells in the
same well. Jurkat cells (a T lymphocyte cell line) show a concentration-dependent increase in internalization of anti-CD3, but not the
two B cell markers. Conversely, Raji cells show a concentration-dependent increase in internalization of anti-CD19 and anti-CD22,
but not anti-CD3. Ramos cells show a concentration-dependent increase in internalization of anti-CD79b, an ADC drug target for
non-Hodgkin's lymphoma.

for multiplexing, no-wash protocols, high-

visualization expedite the process of screening

throughput capabilities, flexibility for a

drug candidates for potential efficacy and

robotic interface, and integrated Forecyt®

toxicity to accelerate antibody discovery and

software for multiparametric data analysis and

development.

14 |

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Sartorius_May21_NeutAntiMakingTheir

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